Identification of Serine-Containing Microcystins by UHPLC-MS/MS Using Thiol and Sulfoxide Derivatizations and Detection of Novel Neutral Losses

Premathilaka, Sanduni H.; Westrick, Judy A.; Isailovic, Dragan

doi:10.1021/acs.analchem.3c04047

Identification of Serine-Containing Microcystins by UHPLC-MS/MS Using Thiol and Sulfoxide Derivatizations and Detection of Novel Neutral Losses

Journal Article · Wed Jan 03 00:00:00 EST 2024 · Analytical Chemistry

DOI:https://doi.org/10.1021/acs.analchem.3c04047· OSTI ID:2583342

Premathilaka, Sanduni H. ^[1]; Westrick, Judy A. ^[2]; ^[1]

Univ. of Toledo, OH (United States)
Wayne State Univ., Detroit, MI (United States)

Microcystins (MCs) are hepatotoxic cyclic heptapeptides produced by cyanobacteria, and their structural diversity has led to the discovery of more than 300 congeners to date. However, with known amino acid combinations, many more MC congeners are theoretically possible, suggesting many remain unidentified. Herein, two novel serine (Ser)-containing MCs were putatively identified in a Lake Erie cyanobacterial harmful algal bloom (cyanoHAB), using high-resolution UHPLC-MS as well as thiol and sulfoxide derivatization procedures. These MCs contain an α,β-unsaturated carbonyl on methyl dehydroalanine (Mdha) residue that undergoes Michael addition to produce a thiol-derivatized MC. Derivatization reactions using various thiolation reagents were followed by MS/MS, and two Python codes were used for data analysis and structural elucidation of MCs. Two novel MCs containing Ser at position 1 (i.e., next to Mdha) were putatively identified as [Ser¹]MC-RR and [Ser¹]MC-YR. Using thiol- and sulfoxide-modified [Ser¹]MCs, identifications were confirmed by the observation of specific neutral losses of the oxidized thiols or sulfoxides in CID-MS/MS spectra in both positive and negative electrospray ionization (ESI) modes. These novel neutral losses are unique for MCs with Mdha and an adjacent Ser residue. In conclusion, data suggest that a gas-phase reaction occurs between oxygen from adjacent Ser residue and sulfur of the Mdha-bonded thiol or sulfoxide, which leads to the formation and detection of stable cyclic MC ions in MS/MS spectra at m/z values corresponding to the loss of oxidized thiols or oxidized sulfoxides from Ser¹-containing MCs.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); US Air Force Office of Scientific Research (AFOSR); USDOE Laboratory Directed Research and Development (LDRD) Program; USDOE National Nuclear Security Administration (NNSA)

Grant/Contract Number:: AC52-07NA27344

OSTI ID:: 2583342

Alternate ID(s):: OSTI ID: 2581395

Report Number(s):: LLNL--JRNL-857791

Journal Information:: Analytical Chemistry, Journal Name: Analytical Chemistry Journal Issue: 2 Vol. 96; ISSN 0003-2700; ISSN 1520-6882

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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